The aim of this study was to investigate the mechanism to reverse the drug resistance of leukemia cells in tetrandrine (Tet) alone or in combination with droloxifen (Drol) by using protein chips and to lay the theoretical basis for the clinical applications. Three monoclonal antibodies against P-glycoprotein (P-gp), the multidrug resistance-associated protein (MRP1) and the breast cancer resistance protein (BCRP) were immobilized onto the agarose gel film-coated glass slides. Protein chips were prepared respectively from K562/A02 cells cultured for 12, 24 and 48 hours with Tet alone or in combination with Drol. The results showed that Tet alone or in combination with Drol could decrease only the expression of P-gp in a time-dependent manner, the effect for 48 hours as follows: Tet + Drol 82.620 +/- 3.227; Tet alone 86.440 +/- 2.906; Drol alone 87.230 +/- 2.049; control 93.670 +/- 2.748 (P < 0.05). However, down-regulation of P-gp by K562/A02 cells cultured with Tet alone or in combination with Drol began at 24 hours (Tet + Drol 85.270 +/- 3.095; control 93.670 +/- 2.748, P < 0.05). The results were coincident with that of FCM. It is concluded that Tet and Drol can downregulate the expression of P-gp in the time-dependent way. There is a significant difference between Tet alone and Tet combined with Drol at 24 hours (P < 0.05). The expression of MRP1 and BCRP are not closely correlated with the reversal mechanism of Tet and Drol, and which may be involved in the mechanism of this combination to reverse multidrug resistance in leukemia.
ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; biosynthesis ; ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; Antineoplastic Agents ; pharmacology ; Benzylisoquinolines ; pharmacology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; drug effects ; Drug Synergism ; Humans ; K562 Cells ; Leukemia ; metabolism ; pathology ; Multidrug Resistance-Associated Proteins ; biosynthesis ; Neoplasm Proteins ; biosynthesis ; Protein Array Analysis ; Tamoxifen ; analogs & derivatives ; pharmacology

OBJECTIVEThe aims of the present study were to assess the characteristics of MSCs which were induced and proliferated in vitro, then to learn the interfaces between them and metal materials.

METHODSAdult dogs were selected as experimental animals, the MSCs (marrow stem cells) were aspirated from the femur, which were induced to the phenotype of OB (osteoblast), then to proliferated in vitro with metal materials. Using biochemical tests, phase contrast microscopy, transmit electromicroscope, scanning electromicroscope to detect MSCs and the image of interface.

RESULTSMSCs had powerful osteogenic phenotype when they were induced in vitro, and they also attached to the metal materials. And there were more cells which attached on tensile stress area than other areas.

CONCLUSIONThe MSCs induced in vitro could be used as source of osteoblast. The affects of air--abrasion with alumina are not significant, but they have relation to the extent of strain and property of stress.

Animals ; Bone Marrow Cells ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Dogs ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Metals ; Osteoblasts

OBJECTIVETo investigate the correlation between visceral adipose tissue-derived serine protease inhibitor (vaspin) concentration and insulin sensitivity in the visceral adipose tissue of young obese Sprague-Dawley (SD) rats.

METHODSTwenty-four SD rats which had been weaned 3 weeks before were randomly divided into two groups (n=12 each) to receive a high-fat and normal diet. The weight and abdominal circumference (AC) of each rat were measured, the fasting plasma glucose (FPG) and fasting insulin (FINS) in blood from the angular vein were measured after 12 hours of fasting and blood glucose (BG) and insulin (INS) levels in blood from the angular vein were measured at 60 and 120 minutes after intraperitoneal injection of 50% glucose (2 g/kg). The rats were sacrificed, and their liver and visceral adipose tissue were weighed. The vaspin concentration of the visceral adipose tissue in each rat was measured using ELISA. Correlation analysis was performed on the vaspin concentration and other indices.

RESULTSCompared with the normal diet group, the high-fat diet group showed significantly higher weight, AC, weight of visceral adipose tissue, FPG, FINS, 120 minute INS level, vaspin concentration, homeostasis model assessment for insulin resistance (HOMA-IR) and homeostasis model assessment of β cell function (HOMA-β) (P<0.05) Insulin sensitivity index (ISI) was significantly lower (P<0.01). Vaspin concentration was positively correlated with visceral adipose tissue and liver weight, AC, 120 minute INS level, FPG, FINS, HOMA-IR and HOMA-β (P<0.05), and negatively correlated with ISI (P<0.05).

CONCLUSIONSHigh expression of vaspin is associated with insulin resistance in young obese SD rats. Vaspin is presumably an adipocytokine that can increase insulin sensitivity, promote insulin secretion by islet β-cells and improve glucose tolerance, and it may be involved in insulin resistance and the disturbance of carbohydrate metabolism.

Animals ; Female ; Glucose Tolerance Test ; Insulin ; blood ; Insulin Resistance ; Intra-Abdominal Fat ; chemistry ; Male ; Obesity ; metabolism ; Rats ; Rats, Sprague-Dawley ; Serpins ; analysis ; physiology

Screening of microorganisms producing flocculating substances was carried out. A strain secreting a large amount of bioflocculant was isolated from wastewater samples collected from the Little Moon River in Beijing. Based on the morphological properties and 16S rDNA sequence analysis, the isolate (designated W31) was classified as Vagococcus sp. A bioflocculant (named MBFW31) produced by W31 was extracted from the culture broth by ethanol precipitation and purified by gel chromatography. MBFW31 was heat-stable and had strong flocculating activity in a wide range of pH with relatively low dosage requirement. MBFW31 was identified as a polysaccharide with molecular weight over 2 x 10(6). It contained neutral sugar and uronic acid as its major and minor components, respectively. Infrared spectra showed the presence of hydroxyl, carboxyl and methoxyl group in its molecules. The present results suggested that MBFW31 had potential application in wastewater treatment.
Carbohydrates ; analysis ; chemistry ; Enterococcus ; genetics ; isolation & purification ; metabolism ; Flocculation ; Species Specificity ; Waste Disposal, Fluid ; methods ; Water Microbiology ; Water Pollutants ; isolation & purification

In order to study the distribution of virulence genes of Listeria monocytogenes (Lm) in Hebei Province,29 virulence genes of Lm,including Listeria monocytogenes pathogenicity islands Ⅰ (LIPI-Ⅰ:prfA,plcA,plcB,hlyA,mpl and actA),10 internalins genes (inlA,inlB,inlC,inlD,inlE,inlF,inlG,inlH /C2,inlI and inlJ) and the other 13 virulence-associated genes (bsh,srtA,iap,sigB,virR,mprF,dltA,dltB,dltC,dltD,srtB,fbpA and hpt) were detected by PCR.Results showed that in the 91 Lm strains,the detection rate of 23 virulence genes were 100％.The 29 virulence genes of 26 Lm strains were all detected,and 65 Lm strains had different deletion of 6 virulence genes inlD,inlF,inlG,inlH /C2,inlJ and mpl.The deletion rate of inlG and inlF were 60.44％ and 54.95％,respectively,following by mpl gene,with a deletion rate of 19.78％.According to the absence of virulence genes,91 strains could be divided into 10 subtypes,and the dominant virulence subtypes was type Ⅰ with all 23 virulence genes.The deletion rate of virulent genes in Shijiazhuang was higher than that in northern Hebei.It is suggested that the rate of virulence gene of food-borne Lm in Hebei Province is high,and the virulence gene deletion patterns has diversity and regional differences.

Objective To discuss the effect of Orem self protection mode on the postoperative rehabilitation knowledge,complications as well as adverse reactions of patients with arteriosclerosis obliterans (ASO) after stent implantation.Methods A total of 100 patients with ASO after stent implantation were randomly divided into the observation group and the control group according to a random number table method,each with 50 cases.The observation group received Orem self-protection mode,while the control group received conventional nursing mode.Two groups rehabilitation knowledge,postoperative complications and adverse reactions were compared.Results There was no statistically significant difference in foot nursing knowledge between two groups (P ＞ 0.05).Patients in the observation group had better knowledge of the disease,walking exercise,Buerger exercise and notes on taking anticoagulant drugs than those in the control group,and the differences were statistically significant (x2 =11.42,9.76,19.05,7.48,respectively; P ＜ 0.01).There were no statistically significant differences in adverse reaction and complications including pulmonary infection,bleeding,cerebral hyperperfusion syndrome and urine retention between two groups (P ＞ 0.05).Cases of lumbago and dorsalgia,pressure sore and re-embolization of lower limb blood vessels were 3,0 and 1 in the observation group,and 13,5 and 6 in the control group,and the differences were statistically significant (x2 =7.44,5.60,3.85,respectively; P ＜0.05).Conclusions Orem self-care mode can promote the postoperative rehabilitation of patients with ASO stent implantation,and improve their self-care ability.

OBJECTIVETo explore the existence of vasculogenic mimicry (VM) in ovarian carcinoma and its correlationship with the clinicopathologic features and prognosis of the tumor.

METHODSA total of 84 ovarian carcinoma cases were collected with complete clinical and prognostic data. CD31 immunohistochemistry and PAS special stain were used to investigate VM in the tumor tissue. Immunohistochemical staining of VEGF, MMP-2, MMP-9, E-cadherin, beta-catenin, and Vimentin were used to explore the pathogenesis of VM.

RESULTSTotally 36 of 84 cases exhibited evidence of VM. FIGO classification, pathologic grades and histological types were significantly different between the VM and non-VM groups. Expression of VEGF, MMP-2, MMP-9, E-cadherin and beta-catenin were higher in the VM group than in the non-VM group. Kaplan-Meier survival curve analysis showed that cases of the VM group had a lower survival rate than that of the non-VM group (P = 0.04).

CONCLUSIONSVasculogenic mimicry exists in ovarian carcinoma. Ovarian carcinomas with a high grade malignancy have a high incidence of VM formation, a higher incidence of metastases and a lower survival rate. High expression of MMP-2 and MMP-9 may contribute to the formation of VM in the ovarian cancer.

Cadherins ; metabolism ; Carcinoma, Endometrioid ; metabolism ; pathology ; Cystadenocarcinoma, Mucinous ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Female ; Humans ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Middle Aged ; Neoplasm Metastasis ; Neovascularization, Pathologic ; metabolism ; pathology ; Ovarian Neoplasms ; metabolism ; pathology ; Survival Rate ; Vascular Endothelial Growth Factor A ; metabolism ; beta Catenin ; metabolism

OBJECTIVETo investigate the effect of simulated microgravity on erythroid differentiation of K562 cells and explore the possible mechanism.

METHODSThe fourth generation rotating cell culture system was used to generate the simulated microgravity environment. Benzidine staining was used to evaluate the cell inhibition rate, and real-time quantitative PCR (qRT-PCR) was used to detect GATA-1, GATA-2, Ets-1, F-actin, β-Tubulin and vimentin mRNA expressions. The changes of cytoskeleton were observed by fluorescence microscopy, and Western blotting was employed to assay F-actin, β-tubulin and vimentin protein expression levels.

RESULTSBenzidine staining showed that simulated microgravity inhibited erythroid differentiation of K562 cells. K562 cells treated with Hemin presented with increased mRNA expression of GATA-1 and reduced GATA-2 and Ets-1 mRNA expressions. Simulated microgravity treatment of the cells resulted in down-regulated GATA-1, F-actin, β-tubulin and vimentin mRNA expressions and up-regulated mRNA expressions of GATA-2 and Ets-1, and reduced F-actin, β-tubulin and vimentin protein expressions. Exposure to simulated microgravity caused decreased fluorescence intensities of cytoskeletal filament F-actin, β-tubulin and vimentin in the cells.

CONCLUSIONSimulated microgravity inhibits erythroid differentiation of K562 cells possibly by causing cytoskeleton damages to result in down-regulation of GATA-1 and up-regulation of GATA-2 and Ets-1 expressions.

Actins ; metabolism ; Cell Differentiation ; Down-Regulation ; GATA1 Transcription Factor ; metabolism ; GATA2 Transcription Factor ; metabolism ; Hemin ; pharmacology ; Humans ; K562 Cells ; Proto-Oncogene Protein c-ets-1 ; metabolism ; Tubulin ; metabolism ; Up-Regulation ; Vimentin ; metabolism ; Weightlessness Simulation

OBJECTIVETo investigate causes and treatment of complications of tracheoesophageal puncture for Blom-Singer voice restoration after total laryngectomy.

METHODSFrom 1986 to 2004, one hundred and fifty one cases with Blom-Singer technique after total laryngectomy for voice restoration were retrospectively analyzed.

RESULTSAll cases were followed up from 6 months to 15 years. Among 151 cases, 138 cases got successful phonation and the total success rate of voice restoration was 91.4%. Of the 151 cases, 15 cases occurred complications. The rate of complications was 9.9%. The common complications included fistula granulations, infection, and leakage. Twelve cases got successful treatment accordingly, and the other 3 cases failed in phonation.

CONCLUSIONSThe procedure of tracheoesophageal puncture for voice restoration is relatively simple and has low complications. this method had high phonation success and good phonation quality, which is one of the best way to make laryngectomee to speak.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Laryngeal Neoplasms ; surgery ; Laryngectomy ; adverse effects ; rehabilitation ; Larynx, Artificial ; Male ; Middle Aged ; Postoperative Complications ; epidemiology ; Retrospective Studies

OBJECTIVETo evaluate the use of protein array chips in detection of multidrug-resistance proteins.

METHODSHuman erythroleukemic cell line K562 and its doxorubicin-resistant counterpart K562/A02 were used in the study. Monoclonal antibodies against P-glycoprotein (P-gP), multidrug resistance-associated protein (MRP1) and breast cancer resistance protein (BCRP) were immobilized onto agarose film-coated glass. The antibody-cell binding was assessed by capturing K562 and K562/A02 cells. The protein array was observed under a microscope and the image was captured with a CCD camera. The expression levels of the three proteins were also measured by flow cytometry (FCM).

RESULTSThe expression of P-gP and BCRP in K562 was very low. However, MRP1 expression was high. P-gP and MRP1 were highly expressed in K562/A02, while the expression of BCRP was low. FCM results showed that the expression rate of P-gP, MRP1 and BCRP in K562 cells was 5.98% +/- 2.19%, 95.80% +/- 3.98%, 1.03% +/- 0.45%, respectively, while that in K562/A02 cells was 92.67% +/- 1.80%, 97.18% +/- 1.02%, 3.98% +/- 0.37%, respectively. The results of protein array method are consistent with those of FCM (P > 0.05).

CONCLUSIONIt is feasible to develop a new protein array technique and to provide a novel method for multi-drug resistant cell detection, with a high throughput, high specificity, simple procedure and low cost.

ATP Binding Cassette Transporter, Sub-Family G, Member 2 ; ATP-Binding Cassette Transporters ; analysis ; ATP-Binding Cassette, Sub-Family B, Member 1 ; analysis ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Humans ; K562 Cells ; Multidrug Resistance-Associated Proteins ; analysis ; Neoplasm Proteins ; analysis ; Protein Array Analysis